• Title/Summary/Keyword: enzyme assay

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Effect of Foliar Spray of Urea on Urease Activity in various Plant Leaves (요소엽면시비(尿素葉面施肥)가 Urease활성(活性)에 미치는 영향(影響))

  • Hong, Jong-Uck;Lee, Hyo-Sa
    • Applied Biological Chemistry
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    • v.24 no.1
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    • pp.15-20
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    • 1981
  • Changes of urease activity in plant leaves following foliar application of urea were investigated with soybean, rye, tomato, radish and cabbage which were actively growing in a field. In this experiment, the procedure of the enzyme assay included incubation of the reaction mixture at $60^{\circ}C$ for 3 hr in order to inactivate heat unstable enzmes which may utilize ammonia produced by urease. The leaves with urea application showed somewhat higher urease specific activities for 2-4 days immediately after the foliar spray as compared with controls. The most difference of the specific activies between urea treatment and control was usually observed 2 days after urea application regardless of the plants. The difference of the specific activities disappeared completely 4 or 5 days following urea treatment. Protein contents in the leaves of soybean and tomato were increased for about 5 days after urea treatment, while no significant difference was found with rye, radish and cabbage. Urea application showed slightly lower ammonia concentration in the leaves which had higher urease activities. These results suggest that foliar spray of urea is very effective when nitrogen supply is required for rapid growth.

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Adverse Reproductive Effects on Plasma Vitellogenin and Sex Steroid Levels, and Gonadosomatic Index in Juvenile Common Carps (Cyprinus carpio) Exposed to 17$\beta$-Estradiol and D-2-Ethylhexyl Phthalate

  • Seo, Jinwon;Park, Kyung-Seo;Moon, Woon-Gi;Lee, Sung-Kyu
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2002.11a
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    • pp.141-144
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    • 2002
  • Environmental estrogens are natural or synthetic substances present in the aquatic environment, especially in effluent from sewage treatment. However, the adverse effects of these estrogenic substances on fish reproduction are unknown. Di-2-ethylhexyl phthalate (DEHP) is the most common phthalate, which Ps used as a plasticizer in polyvinylchloride (PVC), and it is widespread in the environment and has been found in aquatic organisms and sediments. Therefore, juvenile common carps (Cyprinus carpio) were exposed to nominal concentrations of 17$\beta$-estradiol (E2) (0.5, 5, 50 $\mu\textrm{g}$/L) and DEHP (10, 100, 500 $\mu\textrm{g}$/L) for 21 days, to determine the adverse reproductive effects of these compounds on plasma vitellogenin (VTG) induction, sex steroid level, and gonad weight. Electrophoresis (SDS-PAGE) revealed that much of VTG was induced in fish exposed to 5 and 50 E$_2$ $\mu\textrm{g}$/L, but none of DEHP exposure showed induction. Enzyme-linked immunosorbent assay (ELISA) revealed that VTG was significantly induced in fish exposed to 5 and 50 E$_2$ $\mu\textrm{g}$/L, and combination of 50 E$_2$ $\mu\textrm{g}$/L with 10 and 500 DEHP $\mu\textrm{g}$/L, but none of DEHP exposure showed induction. Analysis of sex steroid levels in some fish revealed that testosterone (T) was detected in both male and female fish of the control and DEHP exposures, but none of fish exposed to 22 concentrations had detectable testosterone level. On the other hand, E$_2$ exposure induced 17$\beta$-estradiol in plasma of male fish, but there was no induction of 17$\beta$-estradiol in plasma of male fish exposed to DEHP. Comparison of gonadosomatic index (GSI) revealed that maximal E$_2$ exposure inhibited ovarian growth, but maximal DEHP exposure stimulated testicular growth. The results indicated that those comparisons can be a useful bio-indicator for determining adverse reproductive effect of endocrine disrupting chemicals (EDCs).

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Study on Laboratory Diagnosis of the Ebola Virus and Its Current Trends (에볼라 바이러스 진단법과 개발 동향에 관한 고찰 연구)

  • Jeong, Hye Seon;Kang, Yun-Jung
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.105-111
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    • 2015
  • In late December 2013, the Ebola virus emerged from West Africa. The outbreak started in Guinea and rapidly spread to Liberia and Sierra Leone. Initially, the virus is spread to the human population after contact with infected wildlife and then spread person-to-person through direct contact with body fluids such as blood, sweat, urine, semen, and breast milk. The Ebola virus infects endothelial cells, mononuclear phagocytes and hepatocytes. It causes massive damage to internal tissues and organs, such as blood vessels and the liver, and ultimately death. Most tests for the virus RNA rely on a technology called reverse-transcriptase polymerase chain reaction (RT-PCR). While this method is highly sensitive, it is also expensive, requiring skilled scientists, and delicate power supplies. The strip analytical technique (enzyme-linked immunosorbent assay or ELISA) detects antigens or antibodies to the Ebola virus. This test is cheap and does not require electricity or refrigeration. Despite ongoing efforts directed at experimental treatments and vaccine development, current medical work on the Ebola viral disease is largely limited to supportive therapy. Thus, rapid and reliable diagnoses of the Ebola virus are critically important for patient management, infections, prevention, and control measures.

A Study on the Determinants of Liporotein(a) Level - Primary, Middle, and High School Teachers in Seoul - (혈액내 Lipoprotein(a) 농도의 결정요인에 관한 연구 - 서울시 일부 초, 중, 고교 교사들을 대상으로 -)

  • 최현오;이용욱;나승식;이후장
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.155-163
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    • 1998
  • The purpose of this study was to determine the association between blood lipids, fibrinogen, fasting blood sugar, lifestyle-related factors and lipoprotein(a). This study was performed with 140 healthy adults (male:80, female:60) among the teachers in primary, middle, and high school in Seoul in November 1996. Lipoprotein(a) concentration was measured with an enzyme-linked-immunosorbents assay(ELISA), and the results showed that the distribution was highly skewed and the average concentrations of male and female were $16.94{\pm}12.70\;mg/dL\;and\;17.92{\pm}7.58\;mg/dL$, respectively. The percentage of subject with lipoprotein(a) concentrations higher than 30 mg/dL was 26.4%. Univariate analysis showed significant association between lipoprotein(a) and age, alcohol, meat, and NaCl consumption (p<0.05). In multivariate analysis, lipoprotein(a) correlated positively with age (p<0.05) and meat consumption (p<0.01). These data suggest that lifestyle-related factors may affect the lipoprotein(a) concentration regarded as a risk factor of atherosclerosis disease.

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The Role of Specific IgG in Phagocytosis of Actinobacillus(Haemophilus) Actinomycetemcomitans Y4 by Human Neutrophils (다형핵 백혈구의 Actinobacillus actinomycetemcomitans Y4 균주 탐식시 특이항체의 역할)

  • Kim, Jin-Myung;Chung, Chong-Pyoung;Lee, Young-Hee;Lee, Jin-Yong
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.441-446
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    • 1986
  • Previous studies have demonstrated that phagocytosis of encapsulated bacteria needs the opsonization of bacteria with immunoglobulin and complement. Several investigators have studied the role of specific antibody to the bacteria. The purpose of this study is to investigate the role of specific anti-Actinobacillus actinomycetemcomitans Y4($A{\alpha}Y4$) antibody, which was obtained from the immunized rabbit serum for phagocytosis of $A{\alpha}Y4$ by PMNL. For this study, specific and nonspecific IgG were separated from the sera of the rabbits and PMNL were isolated from 15 healthy adults. By an enzyme-linked immunosorbent assay, the results showed that the binding capacity of anti-$A{\alpha}Y4$ IgG to $A{\alpha}Y4$ was much higher than that of nonspecific IgG; 0.75 and 0.14(O.D. at 400nm), respectively. The oxygen consumption of PMNL, phagocytizing $A{\alpha}Y4$ which was opsonized with specific $A{\alpha}Y4$ IgG(37.13 nmol/min/$1{\times}10^7$ PMNL), was significantly higher than that with nonspecific IgG(27.95 nmol/min/$1{\times}10^7$ PMNL, p<0.01). In immunofluorescence microscopic examination, the difference between the numbers of the ingested $A{\alpha}Y4$ opsonized with specific anti-$A{\alpha}Y4$ IgG and nonspecific IgG reached to statistically significant level; $184{\pm}11.4$ and $133.2{\pm}8.3$ per 100 PMNL, p<0.05. These results suggest that specific anti-$A{\alpha}Y4$ IgG has a significant role in PMNL phagocytosis of encapsulated $A{\alpha}Y4$ and also it can be available to adopt this system to develop anti-capsular antibody to $A{\alpha}Y4$ for enhancing and emphasizing the phagocytic activity against this bacterium.

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Allopurinol Decreases Liver Damage Induced by Dermal Scald Burn Injury (피부 화상으로 유도된 간 손상에서 Allopurinol의 효과)

  • Cho, Hyun-Gug;Yoon, Chong-Guk;Park, Won-Hark
    • Applied Microscopy
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    • v.31 no.1
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    • pp.37-47
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    • 2001
  • In order to investigate a pathogenesis of liver damage induced by skin burn, thermal injury was induced by scald burn on entirely dorsal surface in rats (total burn surface area $20\sim25\%$) except for inhalated injury. At 5 and 24 h after scald burn, biochemical assay and morphological changes in serum and liver tissue were examined. Skin burn increased liver weight (% of body weight, p<0.05) and the activity of serum aniline amino-transferase (ALT, p<0.05), in addition, the activity of xanthine oxidase (XO), an enzyme of oxygen free radical generating system, was elevated (p<0.01) in serum, but not in skin and in liver. Postburn treatment of allopurinol intraperitoneally decreased liver weight, serum ALT activity and serum XO activity. Scald burn induced ultrastructurally swelling of endoplasmic reticulum, ribosome detachment, accumulation of lipid, dilatation of bile canaliculi and intercellular space, neutrophil infiltration, activation of Kupffer's cells and degeneration of hepatocytic microvilli. Futhermore , thermal injury decreased not only the protein concentration in plasma but also the number of intravascular leukocytes, that indicates induction of edema formation with protein exudation and inflammation by neutrophil infiltration into the internal organs. However allopurinol injection after burn inhibited post burn ultrastructural changes. These data suggest that acute dermal scald burn injury leads to liver damage, that is related to elevation of xanthine oxidase activity in serum. Xanthine oxidase may be a key role in the pathogenesis of liver damage induced by skin burn.

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Enhanced Antioxident Effect by over Expression of Tomato β-carotene Hydroxylase Gene (ChyB) Using Agrobacterium-infiltration in Tobacco Plant (Agro-infiltration을 이용한 토마토 β-carotene hydroxylase 유전자(ChyB) 과발현 및 담배식물체의 항산화 효과 증진)

  • Choi, Yoon-Jeong;Yoon, Kyung-Young;Yun, Hae-Keun;Suh, Sang-Gon;Moon, Yong-Sun
    • Horticultural Science & Technology
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    • v.29 no.3
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    • pp.267-272
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    • 2011
  • Several reports indicated that astaxanthin and zeaxanthin have more active anticancer activity than pro-vitamin A carotenes. ${\beta}$-carotene hydroxylase is a key enzyme to synthesize zeaxanthin and astaxanthin in carotenoids biosynthesis pathway. We isolated the ChyB gene encoding ${\beta}$-carotene hydroxylase from tomato leaves. The ChyB gene (1.5Kbp) fragment was cloned into the binary vector and designated to pIG121-ChyB-tom. Agrobacterium-mediated infiltration was used for transient assay in Nicotiana benthamiana. Leaf samples were collected 0, 1, 2, 3 days after infiltration (DAI). RT-PCR result showed that the expression of ${\beta}$-carotene hydroxylase transcripts was not detected in control (0DAI), but its expression was detected after 1 DPI and increased later on. When the activity of ${\beta}$-carotene hydroxylase was measured, the 1,1-diphenyl-pricryl hydrazyl (DPPH) radical scavenging activity (27%) at 2 DAI was significantly higher than that (21%) at 0 DAI. These results indicated that anti-oxidant activity dramatically increased at 2 DAI in tobacco leaves was due to over expression of tomato ${\beta}$-carotene hydroxylase. These results can be the foundation to develop tomato cultivars with high oxy-carotenoids content using the ChyB gene transformation.

Production and Characterization of Monoclonal Antibodies Specific to PAT Protein Expressed in Genetically Modified Herbicide-Resistance Maize (제초제 내성 유전자 변형 옥수수 중 PAT단백질에 특이한 단크론성 항체의 생산과 특성 확인)

  • Kim, Sol-A;Lee, Jeong-Eun;Shim, Won-Bo;Kang, Sung-Jo;Chung, Duck-Hwa
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.193-199
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    • 2018
  • In this study, PAT protein of genetically modified maize was prepared from the recombinant E. coli strain BL21 (DE3), and mice were immunized with the recombinant PAT protein. After cell fusion and cloning, two hybridoma cells (PATmAb-7 and PATmAb-12) were chosen since the monoclonal antibodies (Mabs) produced by them were confirmed to be specific to PAT protein in the indirect enzyme-linked immunsorbent assay (ELISA) and western blot tests. There were no cross-reactions of either Mabs to other GM proteins or to the extracts of non-GM maize. The ELISA based on the PATmAb-7 can sensitively detect 0.3 ng/g PAT protein in corn. These results indicate that the developed Mabs can be used as bio-receptors in the development of immunosensors and biosensors for the rapid and simple detection of GM corn adulterated in foods.

Evaluation of stress-induced analgesia in acupuncture analgesic effect - An Approach on diameters of acupuncture needles and acupuncture point needlings - (침진통 효과에서 스트레스에 의한 진통 효과 개입 여부 평가 - 침굵기, 침자극 부위를 중심으로 -)

  • Jeong, Byeong-Ju;Choe, Il-Hwan;Shin, Hee-Sup;Lim, Sabina
    • Korean Journal of Acupuncture
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    • v.25 no.3
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    • pp.65-80
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    • 2008
  • Objective : In recent years, many investigators have questioned whether the analgesic effect of acupuncture is simply related to the stress-induced analgesia (SIA). However, there has been lack of studies on this issue. In this study, the stress levels induced by manual acupunctures are compared with the stress in animal experiment models. The experiments have been carried out with Sprague Dawley (SD) rats. Method : For stress level evaluation, Hot plate test has been used. Maximum Possible Effect (MPE) has been measured by checking the pre-test time and post-test time. Cortisol and corticosterone concentrations in serum were measured by enzyme-linked immunosorbent assay (ELISA). Results : In the hot plate test, MPE values of post-test time were significantly decreased after 10 minutes than after 5 minutes. Therefore, optimal time interval was chosen as 10 minutes. There was significant difference of MPE values between Suspension group and all other treatment groups. However, there were no significant differences of MPE values between Sham group and all other treatment groups. However, MPE values showed tendency to decrease when acupuncture needle diameter increased. MPE values of ST040, ST040(lido), NAP040(lido) groups were markedly decreased than that of Suspension group, while that of NAP040 group was substantially increased than that of Sham group increased in acupoint and nonacupoint models. Serum cortisol concentrations of treatment groups were not significantly different from that of Suspension and Sham groups. Serum corticosterone concentration of 0.25 mm group was substantially increased than that of compared with Sham group. Serum cortisol and corticosterone concentrations of treatment groups were not significantly different from those of Suspension and Sham groups in acupoint and nonacupoint models. Conclusion : From hot plate test and serum stress hormones concentrations, it is found that manual acupuncture treatment induces negligible stress or SIA on ST36. And the stress induced by manual acupuncture is more closely related to acupuncture point needlings than diameters of acupuncture needles.

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Promoter Cloning of Human SETDB1 Gene Utilizing Bioinformatic Programs (생물정보 프로그램을 활용한 SETDB1 유전자 프로모터 클로닝)

  • Noh, Hee-Jung;Kim, Keun-Cheol
    • Journal of Life Science
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    • v.24 no.1
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    • pp.1-7
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    • 2014
  • Eukaryotic gene expression is an important process, which is initiated by several transcription factors and RNA polymerases that occupy the promoter region of genomic DNA. Although there are many experiments to identify the promoter region in a gene, it is time and labor consuming to finalize it. In this study, we utilized bioinformatic programs, including Ensembl, NCBI, and CpG plots, to identify the cloning promoter region in SETDB1 genomic DNA. We performed PCR amplification to obtain the SETDB1 promoter on an approximately 2 kb region upstream from the TSS named SETDB1-P1. The PCR product was ligated with TA cloning vectors, and we confirmed the insert size using restriction enzyme digestion. Sequentially, the insert was subcloned into a pGL3-luc vector to produce pGL3-SETDB1- P1-luc and then confirmed by DNA sequencing. We also obtained a fragmented PCR product called P2 and P3 and performed a luciferase assay using pGL3-SETDB1-P1-luc transfection. We found that several anticancer drugs, including taxol, 4-FU, and doxorubicin, decreased the promoter activity of SETDB1. We obtained consistent data on the regulation of SETDB1 gene expression after anticancer drug treatment using Western blot analysis and RT-PCR. Our results suggest that promoter cloning of the human SETDB1 gene utilizing bioinformatics is a very useful and timesaving approach to study gene expression.